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摘要：硫酸软骨素是一类具有多种重要生物活性的酸性粘多糖，广泛应用于医药、化妆品和食品等多种领域。通过研究获得产量高、价格便宜、生产周期短及性能优良的硫酸软骨素，对提高它的市场竞争力及拓展其应用领域有着重要的理论意义和现实意义。本文研究的主要内容为：细菌硫酸软骨素类的发酵工艺、细菌硫酸软骨素类发酵工艺条件优化、细菌硫酸软骨素类提取工艺、细菌硫酸软骨素类多糖测定方法的建立等，拟为硫酸软骨素的进一步研究奠定一定的基础。首先，综合单因素实验和正交试验对枯草芽孢杆菌生产硫酸软骨素类的发酵培养基进行了优化，确定了最佳配比如下：蔗糖：2.0%，蛋白胨:2.5%,酵母膏:2.5%，NaCl：0.1%，KH2P04:0.2%,CaCl2:0.05%,MgSO4•7H2O:0.1%。同时，采用单因素实验优化了摇瓶培养的温度、转速、接种量和装液量，优化后的培养基进行摇瓶发酵培养，优化后的培养条件为发酵温度28℃，接种量为4%，转速140rpm，装液量50mL，结合优化后的培养基进行摇瓶发酵，得到硫酸软骨素产量为0.88g/L。同时结合已有的发酵条件，采用5L发酵罐对枯草芽孢杆菌进行扩大发酵，得到硫酸软骨素的产量为1.27g/L。31524
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毕业论文关键字：枯草芽孢杆菌；发酵；优化；提取；测定
Research on fermentation conditions of bacterial chondroitin sulfate
Abstract：Chondroitin sulfate is a kind of acid mucopolysaccharide which has many important biological activity and is widely used in medicine, cosmetic and food, etc. It is of great significance in theory and practice to improve the market competing capacity and expand application area through the research of chondroitin sulfate with high production,inexpensive price,short-cycle and good performance.The main contents of this paper were as following:the fermentation technology of bacterial chondroitin sulfate polysaccharide,the optimization fermentation condition of bacterial chondroitin sulfate polysaccharide , the extraction technology of bacterial chondroitin sulfate , the establishment of the assaying methods of bacterial chondroitin sulfate,which would lay a foundation for further study on chondroitin sulfate.In the fist part of this thesis,by using of single factor experiment and orthogonal experiment,we determined the optimal ratio of the culture medium,that is,sucrose 2.0%，nitrogen sources(peptone-yeast extract) 2.%,，NaCl0.1%,，KH2PO4 0.2%，MgSO4 0.15%。Meanwhile,single factor experiment was applied to optimize shaking culture,including temperature-28℃,rotational speed of shaking tables-160rpm,inocul-m dose-4%and fluid volume-50mL.Based on the optimal medium,we obtained Chondroitin Sulfate with the yield of 0.88g/L.A subsequent 5L-scale fermentation improved the yield to 1.2g/L.
Key words：bacillus subtilis；fermentation；optimization；extraction；assaying
目录
1. 绪论 1
1.1 硫酸软骨素概述 1
1.1.1 硫酸软骨素结构组成 1
1.1.2 硫酸软骨素物理性质 1
1.1.4 硫酸软骨素作用与用途 2
1.2 枯草芽孢杆菌 2
1.3 微生物发酵法 3
1.4 提取方法研究进展 3
1.5 分离纯化 3
1.6 结构分析方法 5
1.7 硫酸软骨素研究的内容及意义 5
2. 材料与方法 6
2.1 菌种 6
2.2 主要试剂与药品 6
2.3主要实验仪器 7
2.4 实验培养基及主要试剂的配制 7
2.5 方法 8
2.5.1 菌种的活化 8
2.5.2 生理生化鉴定 8